فهرست مطالب saeed kaboli

T-cells express two functional forms of the programmed cell death protein 1 (PD-1): membrane (mPD-1) and soluble (sPD-1). The binding of mPD-1 and its ligand (PD-L1) on tumor cells could lead activated lymphocytes toward exhaustion. Selective deletion of the transmembrane domain via alternative splicing of exon-3 in PD-1 mRNA could generate sPD-1. Overexpression of sPD-1 could disrupt the mPD-1/PD-L1 interaction in tumor-specific T cells. We investigated the effect of secreted sPD-1 from pooled engineered and non-engineered T cell supernatant on survival and proliferation of lymphocytes in the tumor microenvironment (TME).

In this experimental study, we designed two sgRNA sequences upstream and downstream of exon-3 in the PDCD1 gene. The lentiCRISPRv2 puro vector was used to clone the dual sgRNAs and produce lentiviral particles to transduce Jurkat T cells. Analysis assays were used to clarify the change in PD-1 expression pattern in the pooled (engineered and non-engineered) Jurkat cells. Co-culture conditions were established with PD-L1+ cancer cells and lymphocytes.

CRISPR/Cas9 could delete exon-3 of the PDCD1 gene in the engineered cells based on the tracking of indels by decomposition (TIDE) and interference of CRISPR edit (ICE) sequencing analysis reports. Our results showed a 12% reduction in mPD-1 positive cell population after CRISPR manipulation and increment in sPD-1 concentration in the supernatant. The increased sPD-1 confirmed its positive effect on proliferation of lymphocytes co-cultured with PDL1+ cancer cells. The survival percent of lymphocytes co-cultured with the pooled cells supernatant was 12.5% more than the control.

The CRISPR/Cas9 exon skipping approach could be used in adoptive cell immunotherapies to change PD-1 expression patterns and overcome exhaustion

During recent years, opium consumption has increased among Iranian diabetic men. The belief that opium consumption can control or even eradicate DM is increasing. This study was conducted to compare the blood sugar, HbA1c, lipid profile and the frequency of diabetes complications in male type II diabetic patient with and without opium consumption. Two hundred thirty two type 2 diabetic men were included in a case- control study between September 2009 and June 2010. They were divided into two groups: with (n= 88), and without (n=144) current opium consumption. Serum glucose was measured by two hexokinase (automized and manual) and orthotolidine method, and Hb A1C was measured by chromatographic ion exchange. The opium consumption was evaluated by thin-layer chromatography (TLC) of urine. Analyses were performed using SPSS and P≤0.05 was considered significant. There was not a significant age difference between two groups. The mean duration of opium use was 36±6 months. The difference between mean serum fasting glucose levels in different methods of measurement was not significant between case and control groups. The results of this study showed a significant lower level of serum HbA1C (p=0.006) and triglyceride (p=0.005) in diabetic patients with opium abuse, but the frequency of peripheral neuropathy and ophthalmic photocoagulation was not different between two groups. Although the HbA1c was lower in males with opium consumption, the frequency of chronic complications of diabetes was not significantly different between two groups. We didnt evaluate other complications of opium consumption, especially mental and physical dependence. In summary, despite observed effects on some of glucose indices, opium is not a harmless treatment for diabetes.